Ovariectomy-induced bone loss in TNFα and IL6 gene knockout mice is regulated by different mechanisms

Abstract

We examined the effects of tumor necrosis factor-α (TNFα) and interleukin-6 (IL6) gene knockout in preserving the bone loss induced by ovariectomy (OVX) and the mechanisms involved in bone metabolism. Twenty female wild-type (WT), -knockout (TNFα) or knockout (IL6) mice aged 12 weeks were sham-operated (SHAM) or subjected to OVX and killed after 4 weeks. Bone mass and skeletal microarchitecture were determined using micro-CT. Bone marrow stromal cells (BMSCs) from all three groups (WT, TNFα and IL6) were induced to differentiate into osteoblasts or osteoclasts and treated with 17-β-estradiol. Bone metabolism was assessed by histological analysis, serum analyses and qRT-PCR. OVX successfully induced a high turnover in all mice, but a repair effect was observed in TNFα and IL6 mice. The ratio of femoral trabecular bone volume to tissue volume, trabecular number and trabecular thickness were significantly decreased in WT mice subjected to OVX, but increased in TNFα mice (1.62, 1.34, 0.27-fold respectively; < 0.01) and IL6 mice (1.34, 0.80, 0.22-fold respectively; < 0.01). Furthermore, we observed a 29.6% increase in the trabecular number in TNFα mice when compared to the IL6 mice. Both, TNFα and IL6 BMSCs exhibited decreased numbers of TRAP-positive cells and an increase in ALP-positive cells, with or without E2 treatment ( < 0.05). While the knockout of or significantly upregulated mRNA expressions of osteoblast-related genes ( and ) and downregulated osteoclast-related mRNA for, and and , knockout appeared to have roles beyond knockout in upregulating mRNA expression and downregulating mRNA expressions of WNT-related genes ( and ) and TNF-related activation-induced genes (). TNFα seemed to be more potentially invasive in inhibiting bone formation and enhancing TRAF6-mediated osteoclastogenesis than IL6, implying that the regulatory mechanisms of TNFα and IL6 in bone metabolism may be different.

Ovariectomy Activates Chronic Low-Grade Inflammation Mediated by Memory T Cells, Which Promotes Osteoporosis in Mice

“…Here, to assess cytokine production, bone marrow CD4 and CD8 were cultured with IL‐7, IL‐15, or both for 48 hours. TNFα and IL‐17A are found in abundance in postmenopausal women with osteoporosis, and TNFα and IL17A null mice do lose bone post‐OVX . Therefore, these two cytokines were assessed in the T cells.…”

Macrophages Switch to an Osteo‐Modulatory Profile Upon RANKL Induction in a Medaka (Oryzias latipes) Osteoporosis Model

“…( 52 ) Also, blocking TNFα by injecting a binding protein or deletion of Tnfa inhibited bone loss induced by ovariectomy. ( 53,54 ) Conversely, TNFα addition promoted osteoclast survival and prevented apoptosis. ( 55 ) Thus, these findings in mice strongly suggest an osteoclast‐promoting effect of TNFα.…”

Bone marrow mesenchymal stem cells: Aging and tissue engineering applications to enhance bone healing

“…However, IL-6, one of the most recognized SASP factors, has been shown to drive osteoclastogenesis [76] and negatively regulate osteoblast differentiation [77]. Knockdown of IL-6 significantly enhanced Runx2 and collagen type I gene expression in osteoblasts while decreasing the expression of osteoclast related genes such as tartrate resistant alkaline phosphatase (TRAP), MMP9, and CTSK [78]. Therefore, it can be extrapolated that SASP production would not be beneficial to bone regeneration.…”